{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,24]],"date-time":"2025-12-24T21:05:41Z","timestamp":1766610341380,"version":"build-2065373602"},"reference-count":23,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2024,1,4]],"date-time":"2024-01-04T00:00:00Z","timestamp":1704326400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Cologne Graduate School of Ageing, CMMC, and CECAD","award":["SFB 1454 P09","851778"],"award-info":[{"award-number":["SFB 1454 P09","851778"]}]},{"name":"Novo Nordisk, Denmark","award":["SFB 1454 P09","851778"],"award-info":[{"award-number":["SFB 1454 P09","851778"]}]},{"name":"European Union\u2019s Horizon 2020 research and innovation program","award":["SFB 1454 P09","851778"],"award-info":[{"award-number":["SFB 1454 P09","851778"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Enteroendocrine cells (EECs) constitute only a small proportion of Villin-1 (Vil1)-expressing intestinal epithelial cells (IECs) of the gastrointestinal tract; yet, in sum, they build the largest endocrine organ of the body, with each of them storing and releasing a distinct set of peptides for the control of feeding behavior, glucose metabolism, and gastrointestinal motility. Like all IEC types, EECs are continuously renewed from intestinal stem cells in the crypt base and terminally differentiate into mature subtypes while moving up the crypt\u2013villus axis. Interestingly, EECs adjust their hormonal secretion according to their migration state as EECs receive altering differentiation signals along the crypt\u2013villus axis and thus undergo functional readaptation. Cell-specific targeting of mature EEC subtypes by specific promoters is challenging because the expression of EEC-derived peptides and their precursors is not limited to EECs but are also found in other organs, such as the brain (e.g., Cck and Sst) as well as in the pancreas (e.g., Sst and Gcg). Here, we describe an intersectional genetic approach that enables cell type-specific targeting of functionally distinct EEC subtypes by combining a newly generated Dre-recombinase expressing mouse line (Vil1-2A-DD-Dre) with multiple existing Cre-recombinase mice and mouse strains with rox and loxP sites flanked stop cassettes for transgene expression. We found that transgene expression in triple-transgenic mice is highly specific in I but not D and L cells in the terminal villi of the small intestine. The targeting of EECs only in terminal villi is due to the integration of a defective 2A separating peptide that, combined with low EEC intrinsic Vil1 expression, restricts our Vil1-2A-DD-Dre mouse line and the intersectional genetic approach described here only applicable for the investigation of mature EEC subpopulations.<\/jats:p>","DOI":"10.3390\/cells13010102","type":"journal-article","created":{"date-parts":[[2024,1,4]],"date-time":"2024-01-04T04:05:38Z","timestamp":1704341138000},"page":"102","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["An Approach to Intersectionally Target Mature Enteroendocrine Cells in the Small Intestine of Mice"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-2774-1164","authenticated-orcid":false,"given":"Christian","family":"Vossen","sequence":"first","affiliation":[{"name":"Obesity and Cancer Research Group, Max Planck Institute for Metabolism Research, Gleueler Strasse 50, 50931 Cologne, Germany"},{"name":"Policlinic for Endocrinology, Diabetes, and Preventive Medicine (PEDP), University Hospital Cologne, 50924 Cologne, Germany"},{"name":"Excellence Cluster on Cellular Stress Responses in Aging Associated Diseases (CECAD), University of Cologne, 50931 Cologne, Germany"},{"name":"Center of Molecular Medicine Cologne (CMMC), University of Cologne, 50931 Cologne, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Patricia","family":"Schmidt","sequence":"additional","affiliation":[{"name":"Obesity and Cancer Research Group, Max Planck Institute for Metabolism Research, Gleueler Strasse 50, 50931 Cologne, Germany"},{"name":"Policlinic for Endocrinology, Diabetes, and Preventive Medicine (PEDP), University Hospital Cologne, 50924 Cologne, Germany"},{"name":"Excellence Cluster on Cellular Stress Responses in Aging Associated Diseases (CECAD), University of Cologne, 50931 Cologne, Germany"},{"name":"Center of Molecular Medicine Cologne (CMMC), University of Cologne, 50931 Cologne, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Claudia Maria","family":"Wunderlich","sequence":"additional","affiliation":[{"name":"Obesity and Cancer Research Group, Max Planck Institute for Metabolism Research, Gleueler Strasse 50, 50931 Cologne, Germany"},{"name":"Policlinic for Endocrinology, Diabetes, and Preventive Medicine (PEDP), University Hospital Cologne, 50924 Cologne, Germany"},{"name":"Excellence Cluster on Cellular Stress Responses in Aging Associated Diseases (CECAD), University of Cologne, 50931 Cologne, Germany"},{"name":"Center of Molecular Medicine Cologne (CMMC), University of Cologne, 50931 Cologne, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Melanie Joyce","family":"Mittenb\u00fchler","sequence":"additional","affiliation":[{"name":"Obesity and Cancer Research Group, Max Planck Institute for Metabolism Research, Gleueler Strasse 50, 50931 Cologne, Germany"},{"name":"Policlinic for Endocrinology, Diabetes, and Preventive Medicine (PEDP), University Hospital Cologne, 50924 Cologne, Germany"},{"name":"Excellence Cluster on Cellular Stress Responses in Aging Associated Diseases (CECAD), University of Cologne, 50931 Cologne, Germany"},{"name":"Center of Molecular Medicine Cologne (CMMC), University of Cologne, 50931 Cologne, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Claas","family":"Tapken","sequence":"additional","affiliation":[{"name":"Obesity and Cancer Research Group, Max Planck Institute for Metabolism Research, Gleueler Strasse 50, 50931 Cologne, Germany"},{"name":"Policlinic for Endocrinology, Diabetes, and Preventive Medicine (PEDP), University Hospital Cologne, 50924 Cologne, Germany"},{"name":"Excellence Cluster on Cellular Stress Responses in Aging Associated Diseases (CECAD), University of Cologne, 50931 Cologne, Germany"},{"name":"Center of Molecular Medicine Cologne (CMMC), University of Cologne, 50931 Cologne, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peter","family":"Wienand","sequence":"additional","affiliation":[{"name":"Obesity and Cancer Research Group, Max Planck Institute for Metabolism Research, Gleueler Strasse 50, 50931 Cologne, Germany"},{"name":"Policlinic for Endocrinology, Diabetes, and Preventive Medicine (PEDP), University Hospital Cologne, 50924 Cologne, Germany"},{"name":"Excellence Cluster on Cellular Stress Responses in Aging Associated Diseases (CECAD), University of Cologne, 50931 Cologne, Germany"},{"name":"Center of Molecular Medicine Cologne (CMMC), University of Cologne, 50931 Cologne, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Paul Nicolas","family":"Mirabella","sequence":"additional","affiliation":[{"name":"Policlinic for Endocrinology, Diabetes, and Preventive Medicine (PEDP), University Hospital Cologne, 50924 Cologne, Germany"},{"name":"Excellence Cluster on Cellular Stress Responses in Aging Associated Diseases (CECAD), University of Cologne, 50931 Cologne, Germany"},{"name":"Research Group Synaptic Transmission in Energy Homeostasis, Max Planck Institute for Metabolism Research, 50931 Cologne, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Leonie","family":"Cabot","sequence":"additional","affiliation":[{"name":"Policlinic for Endocrinology, Diabetes, and Preventive Medicine (PEDP), University Hospital Cologne, 50924 Cologne, Germany"},{"name":"Excellence Cluster on Cellular Stress Responses in Aging Associated Diseases (CECAD), University of Cologne, 50931 Cologne, Germany"},{"name":"Research Group Synaptic Transmission in Energy Homeostasis, Max Planck Institute for Metabolism Research, 50931 Cologne, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7739-486X","authenticated-orcid":false,"given":"Anna-Lena","family":"Schumacher","sequence":"additional","affiliation":[{"name":"FACS & Imaging Core Facility, Max Planck Institute for Biology of Ageing, Joseph-Stelzmann-Str. 9b, 50931 Cologne, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kat","family":"Folz-Donahue","sequence":"additional","affiliation":[{"name":"FACS & Imaging Core Facility, Max Planck Institute for Biology of Ageing, Joseph-Stelzmann-Str. 9b, 50931 Cologne, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1508-0229","authenticated-orcid":false,"given":"Christian","family":"Kukat","sequence":"additional","affiliation":[{"name":"FACS & Imaging Core Facility, Max Planck Institute for Biology of Ageing, Joseph-Stelzmann-Str. 9b, 50931 Cologne, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ingo","family":"Voigt","sequence":"additional","affiliation":[{"name":"Transgenic Core Facility, Max Planck Institute for Biology of Ageing, Joseph-Stelzmann-Str. 9b, 50931 Cologne, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jens C.","family":"Br\u00fcning","sequence":"additional","affiliation":[{"name":"Policlinic for Endocrinology, Diabetes, and Preventive Medicine (PEDP), University Hospital Cologne, 50924 Cologne, Germany"},{"name":"Excellence Cluster on Cellular Stress Responses in Aging Associated Diseases (CECAD), University of Cologne, 50931 Cologne, Germany"},{"name":"Center of Molecular Medicine Cologne (CMMC), University of Cologne, 50931 Cologne, Germany"},{"name":"Department of neuronal Control of Metabolism, Max Planck Institute for Metabolism Research, 50931 Cologne, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Henning","family":"Fenselau","sequence":"additional","affiliation":[{"name":"Policlinic for Endocrinology, Diabetes, and Preventive Medicine (PEDP), University Hospital Cologne, 50924 Cologne, Germany"},{"name":"Excellence Cluster on Cellular Stress Responses in Aging Associated Diseases (CECAD), University of Cologne, 50931 Cologne, Germany"},{"name":"Research Group Synaptic Transmission in Energy Homeostasis, Max Planck Institute for Metabolism Research, 50931 Cologne, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9209-3501","authenticated-orcid":false,"given":"F. Thomas","family":"Wunderlich","sequence":"additional","affiliation":[{"name":"Obesity and Cancer Research Group, Max Planck Institute for Metabolism Research, Gleueler Strasse 50, 50931 Cologne, Germany"},{"name":"Policlinic for Endocrinology, Diabetes, and Preventive Medicine (PEDP), University Hospital Cologne, 50924 Cologne, Germany"},{"name":"Excellence Cluster on Cellular Stress Responses in Aging Associated Diseases (CECAD), University of Cologne, 50931 Cologne, Germany"},{"name":"Center of Molecular Medicine Cologne (CMMC), University of Cologne, 50931 Cologne, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,1,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1003","DOI":"10.1038\/nature06196","article-title":"Identification of Stem Cells in Small Intestine and Colon by Marker Gene Lgr5","volume":"449","author":"Barker","year":"2007","journal-title":"Nature"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2016","DOI":"10.1124\/dmd.114.058404","article-title":"Meta-Analysis of the Turnover of Intestinal Epithelia in Preclinical Animal Species and Humans","volume":"42","author":"Darwich","year":"2014","journal-title":"Drug Metab. 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Rev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"226","DOI":"10.1038\/s41574-019-0168-8","article-title":"Function and Mechanisms of Enteroendocrine Cells and Gut Hormones in Metabolism","volume":"15","author":"Gribble","year":"2019","journal-title":"Nat. Rev. Endocrinol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e74964","DOI":"10.7554\/eLife.74964","article-title":"Enteroendocrine Cell Types That Drive Food Reward and Aversion","volume":"11","author":"Bai","year":"2022","journal-title":"eLife"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"158","DOI":"10.1016\/j.molmet.2019.09.001","article-title":"Single Cell Transcriptomic Profiling of Large Intestinal Enteroendocrine Cells in Mice\u2014Identification of Selective Stimuli for Insulin-like Peptide-5 and Glucagon-like Peptide-1 Co-Expressing Cells","volume":"29","author":"Billing","year":"2019","journal-title":"Mol. 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